Inverse PCR (or alternative) recommendations requested

[Paul N Hengen]

Zunde Wang (U16788 at uicvm.uic.edu) wrote:

> I am considering using inverse PCR to attempt to locate transgenes
> in the mouse genome.

: I am now considering using linker ligations. PanVera sells a PCR
: cloning kit which has six sticky-ended linkers and can be ligated to
: DNA digested with seven restriction enzymes. Since it's quite expensive
: ($350 for 20 reactions), I'd like to ask those who have used it how
: effective the kit is in flanking sequence cloning.

Uhhh...why by a kit for linkers when you can make your own for less?
I suppose it would save you the thought process, but you can do this
without using a kit. Have you considered using vectorettes and a primer
specific to the gene of interest? You only need sequence at one end, right?

@article{Hengen1995Septibs,
author = "P. N. Hengen",
title = "Methods and reagents - Vectorette, splinkerette,
and boomerang {DNA} amplification",
journal = "Trends in Biochemical Sciences",
volume = "20",
number = "9",
pages = "372-373",
month = "sep",
year = "1995"}

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